CONTENTS
Review
- Is the Prion Structure Solved?
PAWEL P. LIBERSKI and JOLANTA BRATOSIEWICZ-ZAPARTAbstract. We report here on the current knowledge on the nature of the scrapie agent or prion. Several lines of evidence suggest that the abnormal isoform of prion protein (PrP) is crucial for scrapie infectivity while evidence that PrP is also a part of the entire particle of the scrapie agent (prion) is much weaker. There is no doubt, however, that conformational changes (transitions from a-helical into ß-pleated structures) of PrP underlay scrapie pathogenesis. In view of the notorious puzzling nature of the scrapie agent, the electron microscopic search for the ultrastructural correlate of it is still warranted. Thus, we discuss the nature of tubulovesicular structures (TVS), the only diseases-specifďc particles known so far and the association between TVS and PrP fibrils which was recently discovered.
Keywords: transmissible spongiform encephalopathies; prion protein (PrP), pathological isoform of PrP; mad cow disease; scrapie.ease; scrapie.
- Immunological Adjuvants and Their Modes of Action.
ANTHONY C. ALLISONAbstract. New adjuvant formulations contain a vehicle, which carries antigens to antigen-presenting cells. Examples of vehicles are liposomes, immune-stimulating complexes and microfluidized squalene-in-water emulsions. Adjuvant formulations may contain immunomodulators, which augment cytokine production, such as a synthetic muramyl dipeptide analog or monophosphoryl lipid A. In a primary cascade of cytokine production at the site of antigen + adjuvant injection, TNF-a promotes the migration of dendritic cells (DC) to lymphoid tissus while GM-CSF accelerates the differentiation of DC into efficient presenters of antigens to T cells. Adjuvants also up-regulate a secondary cascade of cytokines in lymphoid tissues responding to antigenic stimulation: IL-12 augments the production of IFN-y, which favors the production of antibodies of protective isotypes (IgG2a in the mouse). Thus adjuvants can regulate immune responses qualitatively as well as quantitatively. Adjuvant formulations can also activate complement, generating C3d, which binds CD21 on follicular dendritic cells (FDC) and B cells. FDC targeting favors the generation of B lymphocyte memory, which is important for vaccination.
Keywords: adjuvant; dendritic cells; follicular dendritic cells; squalene emulsions; cytokines.okines.
- Molluscan Immune Defenses.
ZDZISLAW GLIŃSKI 1 and JAN JAROSZ 2Abstract. The interest of marine invertebrates as food resources provides a major interest to study molluscan immunity for better understanding of the host response to pathogens. Molluscs possess a natural immunity formed by anatomical and chemical protective barriers that prevent damage of the underlying tissues, body fluid losses and the infections of pathogenic microorganisms and parasites. The main physical barrier is shell and mucus which cover the soft body of molluscs. The integrity of body coverings is supported by blood clotting and wound healing. The internal defense mechanisms of molluscs involve such cellular reactions as: phagocytosis, nodule formation, encapsulation, pearl formation, atrophy, necrosis and tissue liquefaction. Granular hemocytes are the most numerous cell type of molluscan blood active in cellular defenses. Invaders small in size are eliminated by phagocytosis in which participate lectins and products of prophenyloxidase system activation. Numerous and large intruders are eliminated by nodule formation or encapsulation, either cellular or humoral. Humoral components of molluscan immunity are formed by lysozyme activity, lectins and the phenyloxidase system. Up to now the role of mercenenes, paolins, acute phase reactants, a2-macro- globulins and multifunctional binding proteins with anti-protease activity is not well clarified yet. Research prospects on the field of molluscan immunology should essentially be devoted to study cellular defense functions and humoral elfectors to select pathogen-resistant molluscs. This aim could also be achieved by the identification and characterization of immune genes which are candidates for molluscs genetic transformation.
Keywords: molluscan immunity; mucus barriers; encapsulation and nodule formation; pearl formation; paolins; mercenenes.s; mercenenes.
- Rhythms of Immunity.
BARBARA PŁYTYCZ and ROLF SELJELIDAbstract. Rhythms of daily activity are found in all vertebrate species, some of them being diurnal (like humans, dogs, pigeons), others – nocturnal (like mice, rats and bats). Some species undergo very pronounced seasonal changes, as they hibernate in the winter or mate only at the specific seasons. The main regulator (a clock and a calendar) for daily and seasonal rhythms is the periodicity of the external light-darkness, reflected by the periodicity of melatonin secretion from the pineal gland, which is inhibited by light and induced during the darkness. In contrast to melatonin which peaks during the night both in diurnal and noctural species, the cyclicity of other hormones and several immune parameters correlates with the pattern of the animal locomo- tor activity-resting. The immune parameter that peaks at one time of day for a diurnal species peaks about 12 h later for a nocturnal one. Various immune parameters peak at various time points, anticipating an encounter with pathogens during the period of activity while energetically expensive resolution of the immune response during the resting. Daily and seasonal cyclicity of the immune functions are temporally integrated with other physiologic and behavioral processes and all of them are regulated and coordinated with daily and seasonal changes of an external environment by the neuroendocrine homeostatic system.
Keywords: daily rhythms; seasonal rhythms; immunity; immune system; melatonin; adaptation.n; adaptation.
- Regulation of Cytokine Production by Eicosanoids and Nitric Oxide.
JANUSZ MARCINKIEWICZAbstract. Cytokines are widely regarded as regulatory molecules of inflammatory and immune reactions. Nevertheless, the details of functioning of the complex cytokine network are not yet fully understood. Recent data indicate that eicosanoids, primarily the products of the inducible form of cyclooxygenase (COX-2), are involved in the regulation of cytokine production. We have shown that prostaglandins of E series are no longer only suppressor molecules but they selectively up- or down-regulate the cytokine production. Similarly, nitric oxide (NO) generated in activated immune cells by inducible isoform of nitric oxide synthase (iNOS), is considered to be an immunoregulatory molecule. In this article we present a new concept of interactions between cytokines, eicosanoids (prostaglandins and leukotrienes) and NO. Finally, the impact of these molecu- les on the regulation of the immune system is discussed.
Keywords: immunoregulation; cytokines; eicosanoids; prostaglandins; leukotrienes; nitric oxide. oxide.
Clinical Immunology
- Immunological Status of Septic and Trauma Patients. I. High Tumor Necrosis Factor a Serum Levels in Septic and Trauma Patients Are Not Responsible for Increased Mortality; a Prognostic Value of Serum Interleukin 6.
BARBARA ADAMIK, MICHAŁ ZIMECKI, ANDRZEJ WŁASZCZYK and ANDRZEJ KÜBLERAbstract. The aim of this study was to monitor plasma interleukin 6 (IL-6), and tumor necrosis factor a (TNF-a), levels in patients with sepsis, septic shock and multiple organ dysfunction syndrome admitted to the intensive care unit. The patients obtained adequate supportive therapy. Plasma samples were taken upon admission, then on days 1, 2, and 5 following admission. IL-6 and TNF-a levels were determined using bioassays (7TD1 and WEHI-164.13 indicator cell lines, respectively). The results showed that the kinetics of the cytokine release in septic patients differed significantly between survivors and nonsurvivors. In survivors IL-6 concentrations were initially high, fell down rapidly on day 1 after admission, and persisted very low throughout the monitoring time. In contrast, relatively low IL-6 levels in the nonsurvivors, registered upon admission, rose significantly with peak values on day 3 of observation, declining thereafter. TNF-a levels were initially higher in survivors than in nonsurvivors, declined on day 1 following admission, and on day 5 they were higher than the initial values. In nonsurvivors, on the other hand, the starting concentrations of TNF-a were much lower than in survivors with a peak on day 3 with a tendency to fall on day 7. The profiles of cytokine production by traumatic patients (90a/o survivors) revealed low and progressively diminishing levels of IL-6, contrasting with constantly increasing concentratios of TNF-a within the monitoring period. We conclude that high IL-6 levels in septic patients accompanied by high TNF-a levels may indicate bad prognosis. In contrast, rapidly diminishing serum IL-6 levels, even in the presence of high TNF-a levels, could indicate a very good chance for survival. Similar conclusion can be drawn from the monitoring of cytokine production in traumatic, nonseptic patients since almost all of them recovered. We also speculate that TNF-a presence in circulating blood is essential for regeneration of tissues and wound healing.
Keywords: sepsis; trauma; interleukin 6; tumor necrosis factor a.ctor a.
- Modulating Effect of Interleukin 2 Therapy on Interferon Production by Blood Leukocytes of Patients with Minimal Residual Hematological Disease.
MARTYNA KANDEFER-SZERSZEŃ l, WOJCIECH LEGIEĆ 2, ANNA DMOSZYŃSKA2 and AGNIESZKA SZUSTER-CIESIELSKA 1Abstract. This study was designed to investigate the effect of 1.8 x 106 U/day interleukin 2 (IL-2) therapy on interferon (IFN) production. Patients enrolled in the study suffered from multiple myeloma (MM), Hodgkin’s disease (HD) and non-Hodgkin lymphoma (NHL). All of them were in remission after chemotherapy or radiotherapy. Results indicated that IL-2 given subcutaneously at a dose of 1.8 x 106 U/day for 3 weeks induced IFN-y in serum of patients and caused a prolonged effect on the ability of blood leukocytes to produce IFN-y after stimulation in vitro by mitogen phytohemagglutinin (PHA). Such enhancement of IFN-y production may be beneficial for antitumor immune response. Low-dose IL-2 therapy was well tolerated by all patients and side effects not exceeding II grade of toxicity according to WHO scale were observed. Five patients with MM have relapsed 3-10 months after cesation of IL-2 therapy but 15 patients 18 months after tberapy were in complete remission.
Keywords: interleukin 2 low-dose therapy; hematological disease; interferon.e; interferon.
- Isoprinosine and Levamisole as Stimulators of Interferon Production in Blood Leukocytes of Patients with Alcoholic Liver Cirrhosis.
JADWIGA DANILUK 1 and MARTYNA KANDEFER-SZERSZEŃ 2Abstract. Blood leukocytes of 16 patients with alcoholic liver cirrhosis and 18 healthy controls were induced for interferon (IFN) production by phytohemagglutinin (PHA) and concanavalin A (ConA) in the presence or absence of isoprinosine and levamisole at concentrations of 10 pg/ml and 1 ng/ml. This interferon was neutralized in 87-95% by anti-HuIFN-y monoclonal antibodies. In the presence of the drugs the IFN-y production was enhanced, however, IFN-y titers yielded from leukocytes of cirrhotic patients were still below the titers observed in stimulated and unstimulated blood leukocytes of healthy controls. For example, IFN titers induced by PHA in the presence of levamisole (1 ng/ml) in cirrhotic patients were 2.5 times lower (20.2±11.1 U/ml) in comparison to healthy subjects (50.6±27.3 U/ml).
Keywords: isoprinosine; levamisole; interferon y; alcoholic livčr cirrhosis.včr cirrhosis.
- Changes in the Phagocytic Cells in Children Treated with Continuous Ambulatory Peritoneal Dialysis.
MARIA WĄSIK, MARIA BLAIM, DANUTA KOLEWSKA, EWELINA JANOTA-KRAWCZYK, MAŁGORZATA TOMASZEWSKA-PAŃCZYK and MARIA SIENIAWSKAAbstract. Children on continuous ambulatory peritoneal dialysis (CAPD) in endstage renal failure are highly exposed to peritonitis. Peritoneal macrophages (PM) and blood neutrophils (PMNC) are the first line of defense against invading microbes. This study was undertaken for assessing surface receptors expression on PM and PMNC and to check their ability to phagocytosis and killing of bacteria. We have found that in spite of the decreased number of PM in dialysate fluid their viability and activity significantly increased during CAPD. Moreover, higher number of PM expressed CDl6 and CD3S antigens (FcRIII and C3bR, respectively) in comparison with the results observed at CAPD onset. The number of PMNC expressed of these two antigens in uremic children blood were significantly lower in compararison with healthy control. The number of CDl6 positive cells increased under influence of CAPD only temporaly. CAPD caused improvement of phagocytosis and intracellular killing of bacteria by PM but not by PMNC. There is discus- sed here influence of uremia and CAPD on surface antigens, function of phagocytes as well as renewal of PM during CAPD.
Keywords: peritoneal dialysis; phagocytosis; marcophages; blood neutrophils.ophils.
- Eosinophil Cationic Protein in Serum of Corticosteroid-Sensitive and Resistant Bronchial Asthma.
JANUSZ PATKOWSKI, KRZYSZTOF WYTRYCHOWSKI and WOJCIECH MĘDRALAAbstract. Theories for the inflammatory basis of bronchial asthma are presented. The phenomenon of corticosteroid resistance (CR) in bronchial asthma is also discussed. Resistance to corticosteroids, which occurs in about 5% patients with moderate and severe asthma, presents still an important diagnostic and therapeutical problem. In addition, present opinions on the role of eosinophils in the allergic bronchial inflammation were analyzed. The aim of this study was to monitor serum eosinophil cationic protein (ECP) level in asthma patients, sensitive and resistant to glucocorticosteroids (GCS), before and after prednisolone treatment. The resistance to steroids was determined, based on the oral prednisolone test according to Carmichael and vasoconstriction assay according to Stoughton and McKenzie. In the group of cortico- steroid-sensitive (CS) asthmatic patients a statistically significant decrease of ECP level was observed, after 10 day administration of prednisolone in a daily dose of 20 mg, which was associated with a meaningfull increase of FEV, value. On the other hand, the level of ECP in the serum of patients resistant to cortico- steroids, although also decreased under influence of prednisolone, was not correlated with the increase of FEV1 value.
Keywords: corticosteroid-resistant and -sensitive bronchial asthma; eosinophil cationic protein.ionic protein.
- Immunological Status of Patients Subjected to Cardiac Surgery: Effect of Lactoferrin on Proliferation and Production of Interleukin 6 and Tumor Necrosis Factor a by Peripheral Blood Mononuclear Cells in Vitro.
ANDRZEJ WŁASZCZYK, MICHAŁ ZIMECKI, BARBARA ADAMIK, GRAŻYNA DUREK and ANDRZEJ KÜBLERAbstract. The aim of this investigation was to monitor proliferation and cytokine production by peripheral blood mononuclear cells (PBMC) of patients subjected to cardiac surgery. Another goal was to establish regulatory effects of lactoferrin (LF) on these immune reactions in uitro. PBMC were tested before, during surrgery and on day 1 and day 8 -10 following operation. In control donors, low spontaneous and phytohe- magglutinin (PHA)-induced proliferation of PBMC, as well as lipopolysaccharide (LPS)-induced TNF-a secretion was stimulated by LF, but high production of this cytokine was inhibited. In patients, the prolifera- tion of PBMC and the ability to produce IL-6 TNF-a by these cells underwent characteristic changes ;depending on preoperative immune reactivity of patients. In general, low, preoperative reactivity of PBMC showed a tendency to increase within the monitoring period whereas moderate/high responsiveness was diminishing. LF had, in majority of cases, a down-regulatory effect on the proliferative response, best pro- nounced in patients of high/moderate preoperative response. Similarly, LF exhibited, in general, an inhibitory effect on LPS-induced IL-6 production. In terms of TNF-a production, a considerable up-regulatory effect of LF, particularly in low responding patients was of a special interest. In summary, we suggest that LF may play a role in lowering the immune response of patients to surgery and promoting tissue regeneration.
Keywords: cardiac surgery; peripheral blood leukocytes; proliferation; interleukin 6; tumor necrosis factor a; lactoferrin.ferrin.
- Release of Tumor Necrosis Factor a and Its Soluble Receptor p55 in Clinical Septic Syndrome
BOGDAN MODZELEWSKIAbstract. Tumor necrosis factor a (TNF-a, cachectine) attracts much clinical interest when it looses its primary immune properties and, in the septic syndrome, initiates its destructive activity against host tissues. Behaviour of TNF-a and of p55, its soluble receptor (sTNF-R p55), was studied in 50 patients with septic syndrome. Their clinical state was assessed by means of the APACHE II score. Group I consisted of 23 patients who had survived the infection. Twenty seven patients (54%) died, constituting group II. The concentrations of TNF-a and sTNF-R p55 were determined according to the ELISA method. The obtained results were referred to the severity of the patients’ clinical state, their survival rate, the number of lactates and the quantity of the C-reactive protein in blood and the number of blood platelets and neutrophils. It has been documented that in septic syndrome patients with poor prognosis the concentration of TNF-a rises, together with a similar decrease of sTNF-R p55 concentration. This relationship is very important for the prognosis in septic syn- drome.
Keywords: cytokines; tumor necrosis factor; soluble receptor p55; sepsis; septic syndrome.ndrome.
- Distribution of Apolipoprotein E4 Isoform in Alzheimer’s Disease in Poland (Short Communication).
JOLANTA BRATOSIEWICZ-ZAPART, IWONA KŁOSZEWSKA, TOMASZ SOBÓW and PAWEL P. LIBERSKIAbstract. Alzheimer’s disease (AD) is the most common human progressive dementia linked, in its sporadic form, to a common polymorphism within a gene for apolipoprotein E (APOE) mapped to chromosome 19. Individuals with APOE 4 isoforms are more prone to develop sporadic from of AD than those who carry APOE 2 or APOE 3 alleles. As distribution of APOE isoforms in Poland is unknown, we decided to study it in AD and control cases. In AD group, three patients (23.la/o) were homozygous for APOE 4, four cases (30.8%) were heterozygous and six cases (46%) carried no APOE 4 allele. In control group (n-11), only one case (9.1%) was homozygous for APOE 4 allele, no case was heterozygous for APOE 4 while 10 cases (90.9%) carried no APOE 4 alleles. Our results are virtually identical to those reported from Western countries.
Keywords: apolipoprotein E; Alzheimer’s disease.isease.
Immunochemistry
- Haptoglobin Concentration in Blood Sera of Hoofed Mammals in Captivity.
TADEUSZ STEFANIAK, ANIELA GOSPODAREK, FRANZ KLIMA, DIETER SCHADOW and IWONA KĄTNIKAbstract. There were 151 serum samples of the zoological species of hoofed animals from the Bovidae, Cervidae and Equidae families examined for the presence of haptoglobin (Hp). In the majority of samples the Hp levels were close to the ones defined in the related species of domestic animals (86.49, 52.38 and 69.7% in particular families, respectivel . In females (13.94% in the Bovidae family, 63.64% in the Cervidae,15o/o in the Equidae) more often than in males (11.11, 26.32 and Oo/o, respectively) the increased Hp concentrations were found. What is striking it is the relatively high frequency of the raised Hp concentrations occurrence in the Cervidae sera, which may be connected with the higher responsiveness to the factors inducing Hp rise in this family. The contribution of infectious, parasitic, stress-inducing factors and injuries is taken into consideration as the main cause of the raised levels occurrence in the sera of animals kept in zoological gardens. The obtained results suggest the potential usefulness of the Hp estimation as a routine examination in the diagnostics of the intlammation in wild species of Bovidae and Cervidae as well as in monitoring the animal well being in the zoological gardens.
Keywords: haptoglobin; wild Bovidae; wild Cervidae; wild Equidae; zoological garden.garden.
- Some Parameters Influencing Immunoassay of Human and Horse Myoglobins
IWONA E. KOCHANOWSKA, MARIANNA KUROPATWA and APOLINARY SZEWCZUKAbstract. It was noted that human and horse sera as well as human heart and skeletal muscle homogenates or extracts distinctly decrease immunoassays of purified myoglobins. The assays of homogenate and extract myoglobins could be many times increased by precipitation certain proteins with concentrated ammonium sulfate or sodium chloride. Also in homogenates and extracts incubated for several days increased assays of myoglobins were noted. The obtained results indicate that both myoglobins occur in complex with other tissue component(s).
Keywords: human myoglobin; horse myoglobin; immunoassay of myoglobins; polyclonal and monoclonal antibodies.bodies.
- Mechanism of Antigenic Variation in Shigella flexneri Bacilli. IV. Role of Lipopolysaccharides and Their Components in the Sensitivity of Shigella ,flexneri lb and Its Lac+ Recom binant to Killing Action of Serum.
WŁODZIMIERZ DOROSZKIEWICZAbstract. The effect of lipopolysaccharides (LPS) on the normal bovine serum (NBS) bactericidal reactions against mixture of S. flexneri 6713 lb strain and its 3b Lac+ recombinant were investigated. The serum killing of S. flexneri strains was inhibited, in different degree, by LPS extracted from either organisms. These proper- ties were mainly due to LPS molecules; the lipid A fraction showed only low anticomplement activity, the polysaccharide fraction inhibited the killing activity of NBS in very low degree even at high concentration. These studies suggest that LPS composition especially the O-antigen polysaccharide chain contributes to the susceptibility of S. flexneri strains to NBS bactericidal activity.
Keywords: Shigella flexneri; complement; lipid A; lipopolysaccharide.olysaccharide.
Various
- Effect of Viral Infections on the Ability of Human Endothelium to Interferon, Tumor Necrosis Factor and Interleukin 6 Production.
EWA ZACZYŃSKA, ZOFIA BŁACH-OLSZEWSKA, JOLANTA SENDER and WIKTOR JAROSZAbstract. We have reported that cultured human umbilical cord vein endothelial cells (HUVEC) differ from endothelium present on vein surface of organ culture (OC) in production of cytokines and susceptibility to viral infections. In this paper we present the effect of viral infections on interferon (IFN), tumor necrosis factor (TNF), and interleukin 6 (IL-6) production in two culture systems: HUVEC and OC. Infection of 24-48 h HUVEC with herpes simplex type 1 (HSV-1) and vesicular stomatitis virus (VSV) reduced the amounts of IL-6 and TNF produced in comparison to those released spontaneously by uninfected cells. No IFN was detected in media from infected and uninfected HUVEC. Limited viral infections of 3-h-HUVEC and OC usually diminished their efflciency of IL-6 and TNF production. In the case of IL-6 synthesis by OC, effect of viral infection depended, however, on the constitutive synthesis of the cytokine. When spontaneous produc- tion was high ( 800 U/ml), VSV and HSV-1 infection reduced IL-6 level by 2-50 times; in the case of low production (<150 U/ml) the stimulation effect (2-4 fold) was observed. OC released spontaneously some IFN activity (2-32 U/ml). HSV-1 infection of OC reduced IFN level, while VSV in single cases slightly upregulated IFN synthesis.
Keywords: human endothelium; virus infections; interferon; tumor necrosis factor; interleukin 6.ukin 6.
- Metoclopramide and Nitrendipine Are Unable to Enhance Cytotoxicity of Mitoxantrone in Sensitive and Resistant Murine Leukemias in Vitro.
EWA ZACZYŃSKA, ZOFIA BŁACH-OLSZEWSKA, JOLANTA SENDER and WIKTOR JAROSZAbstract. The influence of metoclopramide and nitrendipine on the antitumor action of mitoxantrone (MX) in murine leukemias L1210 and P388 was investigated. These agents were administered in combination with MX to mice bearing sensitive or resistant leukemias. Mitoxantrone resistant P388 was developed ivI tii;o by repeated passages of the tumor to animals treated with suboptimal dose of the drug. Survival time of mice receiving combined therapy were not prolonged as compared with animals treated with MX alone. Both metoclopramide and nitrendipine were unable to overcome MX resistance even partially.
Keywords: mitoxantrone resistance; metoclopramide; nitrendipine; murine leukemias.ine leukemias.
- HIV Proteins-Mediated Alterations in T Cell Interactions with the Extracellular Matrix Proteins and Endothelium (Short Communication).
TOMASZ MROWIEC, CEZARY MELCHAR and ANDRZEJ GÓRSKI
Abstract. Recent data point to the ímmunoregulatory role of extraceIIuIar matríx (ECILf) proteíns ín the process of T cell activation. We studied the effects of HIV proteins on those interactions as well as on endothelial adhesion molecule expression. Gp 120 and gp 41 inhibited CD3-triggered T cell co-stimulation by the ECM proteins, while Nef was stimulatory. T cell adhesion to the ECM proteins was affected in a similar fashion. Furthermore, tat induced E-selectin (but not VCAM-1 and ICAM-1) endothelial expression. Those findings may be relevant for the understanding of immunopathology of AIDS.
Keywords: HIV proteins; endothelium; gp 41; gp 120; Nef; T cells. cells.